The microbiological quality of rainwater used for toilet flushing, cleaning and watering the garden- pilot study 2005

Regenwater opgevangen in reservoirs en toegepast voor onder andere toiletspoeling is vaak fecaal verontreinigd en bevat soms ziekteverwekkende bacterien. Om het infectierisico bij toepassing van dit water te kunnen schatten is aanvullend onderzoek nodig waarbij ziekteverwekkers worden gekwantificeerd en getypeerd en waarbij onderzocht wordt in welke mate gebruikers worden blootgesteld aan het besmette water.Regenwater is aanvankelijk onbesmet, maar bij afstromen langs oppervlakken en tijdens opslag in reservoirs kan besmetting optreden met micro-organismen die ziekte bij de mens kunnen veroorzaken. Dit kan gebeuren wanneer bijvoorbeeld vogelfeces van het dak wordt gespoeld of ratten of andere dieren toegang hebben tot het reservoir of open leidingen. Onderzoek van opgevangen hemelwater op vier verschillende locaties in Nederland toonde de aanwezigheid van de indicatoren voor fecale verontreiniging, bacterien van de coligroep, E. coli en enterococcen, in respectievelijk 28, 27 en 27 van de 28 onderzochte monsters aan. De potentieel ziekteverwekkende bacterien Campylobacter en Legionella pneumophila werden elk een maal op een locatie aangetroffen. Aeromonas en Clostridium perfringens, die ook ziekte bij de mens kunnen veroorzaken, werden in respectievelijk 20 en 23 van de 28 monsters gevonden. Salmonella en Vibrio werden op geen van de locaties aangetroffen. De aanwezigheid van ziekteverwekkende micro-organismen in regenwater toegepast voor toiletspoeling kan negatieve gevolgen voor de volksgezondheid hebben. Op basis van de verkregen resultaten is het nog niet mogelijk om het risico op het oplopen van een infectie bij blootstelling aan dit water te schatten omdat daarvoor nog aanvullende typerings- en blootstellingsgegevens nodig zijn.Rainwater collected in reservoirs and used for toilet flushing, for example, is often fecally contaminated and sometimes contains pathogenic bacteria. Estimating risk of infection caused by use of this water, will require additional research for enumerating and typing pathogens and for determining exposure to the contaminated water. Rainwater at the source is not contaminated with potential human pathogenic micro-organisms, but may become so at surface run-off and during storage in containers. This may occur when bird faeces runs off roofs, or rats or other animals have access to rainwater reservoirs or open mains. Examination of collected rainwater at four different sites in the Netherlands showed the presence of faecal indicator bacteria as total coliforms, E. coli and intestinal enterococci in 28, 27 and 27, respectively, of the 28 samples examined. Each of the potential human pathogenic bacteria, Campylobacter and Legionella pneumophila, was detected once at one sampling site. Aeromonas and Clostridium perfringens, which may also cause disease in humans, were detected in 20 and 23 of the 28 tested samples, respectively. Salmonella and Vibrio were not detected in any of the samples. The presence of potential pathogenic micro-organisms in rainwater used for toilet flushing may have adverse health effects. However, on the basis of the above results, and because additional typing and exposure data are required, it is not yet possible to estimate the risk of infection at exposure to this water.VROM Inspecti

The microbiological quality of rainwater used for toilet flushing, cleaning and watering the garden- pilot study 2005

Rainwater collected in reservoirs and used for toilet flushing, for example, is often fecally contaminated and sometimes contains pathogenic bacteria. Estimating risk of infection caused by use of this water, will require additional research for enumerating and typing pathogens and for determining exposure to the contaminated water. Rainwater at the source is not contaminated with potential human pathogenic micro-organisms, but may become so at surface run-off and during storage in containers. This may occur when bird faeces runs off roofs, or rats or other animals have access to rainwater reservoirs or open mains. Examination of collected rainwater at four different sites in the Netherlands showed the presence of faecal indicator bacteria as total coliforms, E. coli and intestinal enterococci in 28, 27 and 27, respectively, of the 28 samples examined. Each of the potential human pathogenic bacteria, Campylobacter and Legionella pneumophila, was detected once at one sampling site. Aeromonas and Clostridium perfringens, which may also cause disease in humans, were detected in 20 and 23 of the 28 tested samples, respectively. Salmonella and Vibrio were not detected in any of the samples. The presence of potential pathogenic micro-organisms in rainwater used for toilet flushing may have adverse health effects. However, on the basis of the above results, and because additional typing and exposure data are required, it is not yet possible to estimate the risk of infection at exposure to this water.Regenwater opgevangen in reservoirs en toegepast voor onder andere toiletspoeling is vaak fecaal verontreinigd en bevat soms ziekteverwekkende bacterien. Om het infectierisico bij toepassing van dit water te kunnen schatten is aanvullend onderzoek nodig waarbij ziekteverwekkers worden gekwantificeerd en getypeerd en waarbij onderzocht wordt in welke mate gebruikers worden blootgesteld aan het besmette water.Regenwater is aanvankelijk onbesmet, maar bij afstromen langs oppervlakken en tijdens opslag in reservoirs kan besmetting optreden met micro-organismen die ziekte bij de mens kunnen veroorzaken. Dit kan gebeuren wanneer bijvoorbeeld vogelfeces van het dak wordt gespoeld of ratten of andere dieren toegang hebben tot het reservoir of open leidingen. Onderzoek van opgevangen hemelwater op vier verschillende locaties in Nederland toonde de aanwezigheid van de indicatoren voor fecale verontreiniging, bacterien van de coligroep, E. coli en enterococcen, in respectievelijk 28, 27 en 27 van de 28 onderzochte monsters aan. De potentieel ziekteverwekkende bacterien Campylobacter en Legionella pneumophila werden elk een maal op een locatie aangetroffen. Aeromonas en Clostridium perfringens, die ook ziekte bij de mens kunnen veroorzaken, werden in respectievelijk 20 en 23 van de 28 monsters gevonden. Salmonella en Vibrio werden op geen van de locaties aangetroffen. De aanwezigheid van ziekteverwekkende micro-organismen in regenwater toegepast voor toiletspoeling kan negatieve gevolgen voor de volksgezondheid hebben. Op basis van de verkregen resultaten is het nog niet mogelijk om het risico op het oplopen van een infectie bij blootstelling aan dit water te schatten omdat daarvoor nog aanvullende typerings- en blootstellingsgegevens nodig zijn

Prevalence and determinants of HPV infection among Colombian women with normal cytology

Human papillomavirus is the principal risk factor associated with cervical cancer, the most common malignancy among women in Colombia. We conducted a survey, aiming to report type specific prevalence and determinants of human papillomavirus infection in women with normal cytology. A total of 1859 women from Bogota, Colombia were interviewed and tested for human papillomavirus using a general primer GP5+/GP6+ mediated PCR–EIA. The overall HPV DNA prevalence was 14.8%; 9% of the women were infected by high risk types, 3.1% by low risk types, 2.3% by both high risk/low risk types and 0.4% by uncharacterized types (human papillomavirus X). Thirty-two different human papillomavirus types were detected, being human papillomavirus 16, 58, 56, 81(CP8304) and 18 the most common types. The human papillomavirus prevalence was 26.1% among women younger than 20 years, 2.3% in women aged 45–54 years, and 13.2% in women aged 55 years or more. For low risk types the highest peak of prevalence was observed in women aged 55 years or more. Compared to women aged 35–44 years, women aged less than 20 years had a 10-fold increased risk of having multiple infections. Besides age, there was a positive association between the risk of human papillomavirus infection and number of regular sexual partners and oral contraceptive use. In women aged below 25 years, high educational level and having had casual sexual partners predicted infection risk. In conclusion, there was a broad diversity of human papillomavirus infections with high risk types being the most common types detected. In this population multiplicity of sexual partners and, among young women, high educational level and casual sexual partners seem to determine risk

Touchdown General Primer (GP5+/GP6+) PCR and optimized sample DNA concentration support the sensitive detection of human papillomavirus

BACKGROUND: The GP5+/GP6+ PCR assay is a well-established HPV detection technique. This study has examined the effects of incorporating 'hot start' and 'touchdown' steps into the protocol. In addition, dTTP was substituted with dUTP to permit contamination control measures against carry-over PCR product. METHODS: Firstly, HPV-16 was amplified from SiHa cell DNA (0.1 ng–100 ng) diluted in a background of C-33A DNA (100 ng-2 μg). Secondly, the detection of small quantities (15ag-1.5pg) of HPV recombinant plasmids (types 16, 31, 33, 45, 51, 52, and 56) diluted in C-33A DNA was investigated. Thirdly, clinical sample DNA extracts (cervical smears, formalin-fixed vaginal lesions and breast tumors) were tested for HPV. Six different PCR protocols were assessed. HPV was detected by gel electrophoresis, and by Southern and dot blot hybridization. RESULTS: HPV detection sensitivity was dependent on the total amount of DNA in a PCR. Touchdown protocols supported HPV-16 detection from 1 ng or 0.5 ng SiHa cell DNA in a background of 2 μg or 1 μg C-33A DNA respectively, and from 0.1 ng of SiHa cell DNA (~28 copies HPV-16) in 500 ng or 100 ng background DNA. Under standard GP5+/GP6+ annealing conditions, HPV-16 went undetected when the DNA content of a PCR was 2 μg or 1 μg, and with 500 ng C-33A DNA the sensitivity limit was 1 ng SiHa cell DNA. HPV recombinant plasmids were each detected with high (albeit varying) sensitivity by a touchdown protocol. HPV-31 was better amplified under standard annealing conditions (1.5fg in 100 ng background DNA) than by a touchdown approach (15fg detection limit). HPV-52 was not amplified by the standard protocol at the dilutions tested. Seventeen different HPV types were demonstrated in 47/65 (72%) abnormal cytology samples recorded as HPV negative by standard GP5+/GP6+ conditions. Twenty-one different HPV types were recorded in 111/114 (97%) vaginal lesions. Multiple infections were also detectable using a touchdown approach. Of 26 breast tumors, 5 (19%) tested HPV positive by the standard assay and 15/26 (58%) using a touchdown protocol. CONCLUSION: Touchdown modification of the GP5+/GP6+ PCR assay enables the detection of HPV undetected under regular assay conditions. The use of standardized DNA quantities in a PCR rather than standard sample volumes containing arbitrary amounts of DNA is supported. A touchdown approach may be beneficial as an analytical test for the re-evaluation of (apparently) HPV negative abnormal cervical cytological or histological samples, and for investigating the association of HPV with disease conditions at diverse organ sites. The clinical utility of a touchdown approach for HPV detection requires further investigation as increased assay analytical sensitivity may not necessarily equate with improved clinical sensitivity or specificity

Bronnen van antibioticaresistentie in het milieu en mogelijke maatregelen

Het RIVM heeft in samenwerking met andere instituten onderzocht hoeveel resistente bacteriën via afvalwater in het Nederlandse oppervlaktewater terechtkomen. Afvalwater is, naast mest, de belangrijkste bron waardoor antibioticaresistente bacteriën in het milieu belanden. In 60 tot 100% van het onderzochte afvalwater zitten bijzonder resistente micro-organismen(BMRO), zoals ESBL-producerende E. coli en carbapenemresistente Enterobacteriaceae. Daarnaast zijn resten van antibiotica in het afvalwater gevonden. Mensen kunnen aan resistente bacteriën in het milieu worden blootgesteld, bijvoorbeeld als zij in contact komen met water waarop gezuiverd afvalwater wordt geloosd. Vooralsnog is het onduidelijk hoe groot de bijdrage van deze blootstelling is ten opzichte van andere blootstellingsroutes, en wat de gevolgen daarvan voor de volksgezondheid zijn. Daarvoor wordt aanvullend onderzoek aanbevolen. Duidelijk is wel dat mensen naast blootstelling via het milieu, ook door andere bronnen kunnen worden blootgesteld aan antibioticaresistente bacteriën, bijvoorbeeld tijdens reizen naar landen in Azië of Zuid-Amerika. Door de huidige behandeling van afvalwater nemen de concentraties van antibioticaresistente bacteriën af. Er bestaan aanvullende zuiveringstechnieken voor afvalwater die het aantal antibioticaresistente bacteriën in het oppervlaktewater nog verder kunnen verminderen. Ook de concentraties van resistente bacteriën in mest kunnen door aanvullende zuiveringstechnieken verminderd worden.In collaboration with other institu tions , RIVM investigated how many resistant bacteria enter Dutch surface water s from waste water treatment plants. Next to manure, waste water is the main source of resistant bacteria in the environment. Resistant bacteria of particular public health concern, including ESBL -producing E. coli and carbapenem resistant Enterobacteriace ae, have been found in 60 -100% of all investigated waste water samples. Antibiotics themselves have also been found in waste water. Human exposure to resistant bacteria is possible, for example, upon contact with surface water which receives waste water treatment plant effluent. So far, the contribution of this exposure to the total exposure to resistant bacteria is unknown, as are the effects of this exposure on public health. Additional research is therefore recommended. Humans can also be exposed to resistant bacteria via other routes, such as travel to Asian or Latin American. The current treatment of waste water reduces the concen trations of resistant bacteria. Advanced techniques are available which can decrease the amounts of resistant bacteria further. Options for the reduction of resistant bacteria also exist for the treatment of manure

High-risk human papillomavirus clearance in pregnant women: trends for lower clearance during pregnancy with a catch-up postpartum

We followed 353 women referred with abnormal cervical cytology in a non-intervention cohort study. In 91 pregnant women we compared high-risk human papilloma virus rates in the subsequent trimesters and postpartum in comparison to 262 non-pregnant women. High-risk human papilloma virus clearance was compared with 179 high-risk human papilloma virus positive non-pregnant women. Our main questions were: (1) do high-risk human papilloma virus rates change during pregnancy?; and (2) is there any difference between high-risk human papilloma virus clearance in pregnant and non-pregnant women? Women were monitored 3–4 monthly by cytology, colposcopy, and high-risk human papilloma virus testing. The median follow-up time was 33 months (range 3–74). Non-pregnant women showed prevalence rates of high-risk human papilloma virus of 64, 57, 53, and 50%, respectively, in four subsequent 3-months periods since the start of the study. These high-risk human papilloma virus rates were higher than in the three trimesters of pregnancy, and during the first 3 months postpartum, i.e. 50, 44, 45, and 31%, respectively. Postpartum only, this difference was statistically significant (P=0.004). Paired comparisons of high-risk human papilloma virus prevalence rates of the different trimesters with the postpartum rate showed (McNemar test) decreased rates: first trimester: 18% (P=0.02), second trimester: 13% (P=0.02) and third trimester: 23% (P<0.005). Such a phenomenon was not found in non-pregnant women. Pregnant women showed a trend for increased high-risk human papilloma virus clearance during the third trimester and postpartum compared to non-pregnant women (hazard ratios 3.3 (0.8–13.7) and 4.6 (1.6–12.8), respectively). These results suggest a lowered immune-response against human papilloma virus during the first two trimesters of pregnancy with a catch-up postpartum

Prevalence of type-specific HPV infection by age and grade of cervical cytology: data from the ARTISTIC trial

Human papillomavirus (HPV) infection causes cervical cancer and premalignant dysplasia. Type-specific HPV prevalence data provide a basis for assessing the impact of HPV vaccination programmes on cervical cytology. We report high-risk HPV (HR-HPV) type-specific prevalence data in relation to cervical cytology for 24 510 women (age range: 20–64; mean age 40.2 years) recruited into the ARTISTIC trial, which is being conducted within the routine NHS Cervical Screening Programme in Greater Manchester. The most common HR-HPV types were HPV16, 18, 31, 51 and 52, which accounted for 60% of all HR-HPV types detected. There was a marked decline in the prevalence of HR-HPV infection with age, but the proportion due to each HPV type did not vary greatly with age. Multiple infections were common below the age of 30 years but less so between age 30 and 64 years. Catch-up vaccination of this sexually active cohort would be expected to reduce the number of women with moderate or worse cytology by 45%, but the number with borderline or mild cytology would fall by only 7%, giving an overall reduction of 12% in the number of women with abnormal cytology and 27% in the number with any HR-HPV infection. In the absence of broader cross-protection, the large majority of low-grade and many high-grade abnormalities may still occur in sexually active vaccinated women

Human Papillomavirus (HPV) in breast tumors: prevalence in a group of Mexican patients

<p>Abstract</p> <p>Background</p> <p>Breast cancer is one of the main health problems in developed countries, occupying first place in mortality in women. It is well-known that there are risk factors associated with breast cancer development. Nonetheless, in 50–80% of cases known risk factors have not been identified, this has generated the attempt to identify new factors related with this neoplasia as viral infections. The aim of this work is investigate the prevalence of HPV DNA in patients with breast lesions at the Instituto Nacional de Cancerologia de Mexico.</p> <p>Methods</p> <p>Fifty-one cases of breast cancer were selected from the files of the institute and compared by age and tumor size with 43 cases of non malignant breast lesions (fibroadenoma, fibrocystic disease and phyllodes tumor). Paraffin embedded specimens were selected, HPV DNA was analyzed by polymerase chain reaction (PCR) and sequenced for different types of HPV in case of positivity for HPV-DNA. Descriptive analysis of clinical and pathological variables was performed and comparisons between positive and negative cases was done.</p> <p>Results</p> <p>All patients were mexican, mean age was 53.3, median age of menarche was 13 and median tumor size 9 cms. Cervicovaginal cytology was performed to all patients, 1 patient (1.9%) of cancer group had HPV and none in the other group, no cases were diagnosed with cervical dysplasia. In the group of carcinomas 36 (70.5%) were negative and 15 (29.4%) were positive to HPV-DNA, 10(66.6%) were positive for HPV 16, 3(20%) for HPV 18, two cases (13.4%) were positive for both. In the group of benign conditions all were negative to HPV-DNA.</p> <p>Conclusion</p> <p>Presence of HPV in breast cancer in our group of cases is high in comparison to other authors; larger numbers of cases need to be analyzed in order to establish the exact role of this virus in the pathogenesis of breast cancer.</p

A quantitative polymerase chain reaction-enzyme immunoassay for accurate measurements of human papillomavirus type 16 DNA levels in cervical scrapings

A quantitative polymerase chain reaction-enzyme immunoassay (Q-PCR-EIA) was developed to measure the amount of human papillomavirus (HPV) 16 DNA per genome equivalent in cervical scrapings. The quantitative approach was based on a combined competitive PCR for both HPV 16, using the general primer GP5+/6+ PCR, and β-globin DNA. The two competitive PCRs involve co-amplification of target sequences and exogenously added DNA constructs carrying a rearranged 30 bp sequence in the probe-binding region. The accuracy of quantification by combining the two competitive PCR assays was validated on mixtures of HPV 16 containing cervical cancer cells of CaSki and SiHa cell lines. Comparison of this fully quantitative PCR assay with two semi-quantitative HPV PCR assays on a series of crude cell suspensions from HPV 16 containing cervical scrapings revealed remarkable differences in the calculated relative HPV load between samples. We found evidence that correction for both intertube variations in PCR efficiency and number of input cells/integrity of DNA significantly influence the outcome of studies on viral DNA load in crude cell suspensions of cervical scrapings. Therefore, accurate measurements on viral DNA load in cervical scrapings require corrections for these phenomena, which can be achieved by application of this fully quantitative approach. © 1999 Cancer Research Campaig